Background: Amoebiasis, or amoebic dysentery, is an infection caused by protozoan Entamoeba histolytica of humans that causes 4000-100,000 deaths annually. Recommendation treatment of amoebiasis using metronidazole have been reported to be less effective, due to the drug resistance effect on Entamoeba histolytica. Previous research has reported that the dichloromethane extract of leaves and the dichloromethane extract of stem from Garcinia parviflora inhibit amoeba cell with IC₅₀value of 38.59 ± 9.46 µg/mL and 68.34 ± 0.4 µg/mL, respectively. Objective: This research was designed to determine the activity of the Garcinia parviflora fractions from dichloromethane leaves extract against Entamoeba histolytica. Materials and Methods: Dichloromethane extract of leaves has been fractionated by Vacuum Liquid Chromatography (VLC) to obtain seven fractions. Their anti-amoebic activity were determined by in vitro cell-based assay against Entamoeba histolytica HM-1:IMSS (clone 6) strain and CS3 enzymatic assay. Result: Cell based and CS3 Enzymatic assay showed that out of seven fractions, fraction 2 had the highest anti-amoebic activity with inhibitory concentration (IC₅₀) of 16.13 µg/mL and 42.24 µg/mL, respectively. Conclusion: Based on the data, the fraction 2 of dichloromethane extract of Garcinia parviflora leaves was the most active and could be a potential candidate in the development of anti-amoebic drugs through cell based and CS3 enzymatic assay.